摘要
基于ITS2序列和HPLC指纹图谱建立琴叶榕及其近缘种的鉴别方法,为琴叶榕质量控制提供参考。采用MEGA6.0软件分析ITS2序列,计算K2P遗传距离,构建NJ树,ITS2 database在线分析二级结构。采用HPLC建立指纹图谱并测定补骨脂素和佛手柑内酯含量。琴叶榕ITS2序列长度210 bp,平均GC含量62.37%;近缘种的ITS2序列长度范围为218~232 bp,GC含量范围为60.25%~64.03%;琴叶榕种内K2P平均遗传距离小于琴叶榕与近缘种种间的平均K2P遗传距离;NJ树可将琴叶榕及其近缘种区分,且ITS2二级结构的臂数目、臂长度、臂上茎环数目、臂与臂的夹角均有明显差异。不同产地琴叶榕的HPLC指纹图谱相似度为0.902~0.981,确定10个共有峰,8号峰为补骨脂素、9号峰为佛手柑内酯,含量分别为1.01~4.80、0.96~4.88 mg/g;近缘种与琴叶榕对照图谱的相似度均小于0.807,全缘琴叶榕、条叶榕、粗叶榕的补骨脂素含量分别为0.38~0.63、1.08~1.32、2.45~14.73 mg/g,条叶榕、粗叶榕的佛手柑内酯含量分别为0.25~0.34、0.13~0.74 mg/g,全缘琴叶榕未检出佛手柑内酯。结果表明ITS2序列和HPLC指纹图谱可用于鉴定琴叶榕与近缘种,建立的HPLC指纹图谱简便,专属性强,重现性良好,为琴叶榕的质量评价提供了参考。
TS2 sequences and high-performance liquid chromatography(HPLC)fingerprint method were established to provide reference for the quality control of F.pandurata.MEGA6.0 was used to analyse ITS2 sequences,calculate K2P genetic distances,construct neighbor joining(NJ)phylogenetic tree.Secondary structure was analyzed by online ITS2 database.HPLC was used to establish the fingerprint and determine the content of psoralen and bergapten.The length of ITS2 sequence from F.pandurata is 210 BP and the average GC content is 62.37%.The length of ITS2 sequence of the related species ranged from 218 to 232 BP,and the content of GC ranged from 60.25%to 64.03%.The average genetic distance of K2P from F.pandurata was less than the interspecific genetic distance between F.pandurata and its related species.NJ tree showed that ITS2 could distinguish F.pandurata and its related species,and there were significant differences in the number of arms,arm length,number of stem rings on arms and the angle between arms in ITS2 secondary structure.The similarity of HPLC fingerprints from F.pandurata in different habitats ranged from 0.902-0.981 and the data showed 10 common peaks.Peak 8 was identified as psoralen,peak 9 was bergapten,and the content of psoralen was 1.01-4.80 mg/g,bergapten was 0.96-4.88 mg/g.The similarity of the common pattern between the related species and F.pandurata was less than 0.807.The content of psoralen in F.pandurata var.holophylla,F.pandurata var.angustifolia,and F.hirta was 0.38-0.63,1.08-1.32,2.45-14.73 mg/g,respectively.The content of bergapten in F.pandurata var.angustifolia was 0.25-0.34,and F.hirta was 0.13-0.74 mg/g.The content of bergapten was not detected in F.pandurata var.holophylla.The results showed that ITS2 sequences and HPLC fingerprint can be used for identification of F.pandurata and its related species.The established HPLC fingerprint analysis method is simple,specificity and reproducibility,which provides a reference for the quality evaluation of F.pandurata.
作者
张声源
庄远杯
杨秋娜
魏爱红
张超
杨亚利
ZHANG Sheng-yuan;ZHUANG Yuan-bei;YANG Qiu-na;WEI Ai-hong;ZHANG Chao;YANG Ya-li(Guangdong Provincial Key Laboratory of Conservation and Precision Utilization of Characteristic Agricultural Resources in Mountainous Areas,Meizhou 514015,China;Institute of Hakka Medicinal Bio-resources,Medical College,Jiaying University,Meizhou 514031,China)
出处
《天然产物研究与开发》
CAS
CSCD
2023年第4期640-647,676,共9页
Natural Product Research and Development
基金
广东省中医药局项目(20212243)
梅州市社会发展科技计划(2020B151)
2022年广东省基地嘉应学院客家研究院大客家平台研究团队(22KYKT03)
广东省大学生创新创业项目(S202010582082)。
关键词
琴叶榕
ITS2序列
HPLC指纹图谱
近缘种
Ficus pandurata Hance
ITS2 sequences
high-performance liquid chromatography fingerprint
related species
